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1.
J Biol Regul Homeost Agents ; 33(3 Suppl. 1): 11-17, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31538445

RESUMO

In vivo Reflectance Confocal Microscopy (RCM) allows to optically biopsy vital tissues, non-invasively and in real time. It results in horizontal virtual slices at a microscopic resolution and correlating with conventional histopathology. The aim of the present work is to describe RCM cellular and architectural findings in oral mucosae affected by erosive-ulcerative diseases, thus highlighting in vivo the wellknown histological peculiarities. A series of conventionally diagnosed Recurrent Aphthous stomatitis (RAS) and Pemphigus Vulgaris (PV) erosive and/or ulcerative oral lesions underwent RCM imaging to establish the application of RCM imaging to this kind of inflammatory non-tumoral lesions. A total of 12 RAS-related lesions and 8 PV-related lesions were considered. RCM imaging was capable to visualize their microscopic peculiarities, mainly inflammatory infiltrate, vessel dilation (RAS) and acantholytic cells, intraepithelial clefts and inflammatory cell carpets (PV). Despite RCM may result unnecessary to diagnose oral lesions referred to RAS and PV, its capability to highlight their main microscopic features could be advantageously used to monitor the healing or worsening of the clinical situation as well as the responsiveness/refractoriness to therapy.


Assuntos
Microscopia Confocal , Mucosa Bucal/diagnóstico por imagem , Mucosa Bucal/patologia , Pênfigo/diagnóstico por imagem , Estomatite Aftosa/diagnóstico por imagem , Biópsia , Humanos
2.
J Biol Regul Homeost Agents ; 33(3 Suppl. 1): 19-25, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31538446

RESUMO

Oral mucosa pigmentations belong to a heterogeneous variety of lesions, which are usually divided into two groups: exogenous or endogenous pigmentations. The pigmented lesions most frequently found in the oral mucosa are the amalgam tattoo, the melanotic macula and the nevus. All these lesions may affect every part of the oral mucosa, and they may represent a hard diagnostic challenge for the clinician; the clinical objective examination is not sufficient to make a correct diagnosis. Reflectance Confocal Microscopy provides a real-time microscopic evaluation of tissue layers, and is widely considered a useful auxiliary tool in monitoring skin and mucosa lesions. In this context, Reflectance Confocal Microscopy imaging is a valid aid in the management of oral mucosa pigmented lesions, to corroborate and support the diagnostic process.


Assuntos
Lábio/diagnóstico por imagem , Microscopia Confocal , Nevo Pigmentado/diagnóstico por imagem , Pigmentação , Humanos , Neoplasias Cutâneas/diagnóstico por imagem
3.
Exp Parasitol ; 205: 107748, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31442453

RESUMO

Trypanosoma cruzi (the causative agent of Chagas disease) presents a complex life cycle that involves adaptations in vertebrate and invertebrate hosts. As a protozoan parasite of hematophagous insects and mammalian hosts, T. cruzi is exposed to reactive oxygen species (ROS). To investigate the functionality of T. cruzi tartrate-resistant acid phosphatase type 5 (TcACP5), we cloned, superexpressed and purified the enzyme. Purified TcACP5 exhibited a Vmax and apparent Km for pNPP hydrolysis of 7.7 ±â€¯0.2 nmol pNP × µg-1 × h-1 and 169.3 ±â€¯22.6 µM, respectively. The pH dependence was characterized by sharp maximal activity at pH 5.0, and inhibition assays demonstrated its sensitivity to acid phosphatase inhibitors. Similar activities were obtained with saturating concentrations of P-Ser and P-Thr as substrates. The enzyme metabolizes hydrogen peroxide (H2O2) in vitro, and parasites superexpressing this enzyme were more resistant to oxidative stress promoted by H2O2. Taken together, these results suggest that TcACP5 plays a central role in phosphoryl transfer and redox reactions.


Assuntos
Peróxido de Hidrogênio/farmacologia , Estresse Oxidativo/fisiologia , Fosfatase Ácida Resistente a Tartarato/metabolismo , Trypanosoma cruzi/enzimologia , Sequência de Aminoácidos , Imunofluorescência , Regulação Enzimológica da Expressão Gênica , Peróxido de Hidrogênio/metabolismo , Concentração de Íons de Hidrogênio , Microscopia Confocal , Oxirredução , Especificidade por Substrato , Fosfatase Ácida Resistente a Tartarato/antagonistas & inibidores , Fosfatase Ácida Resistente a Tartarato/química , Transfecção , Trypanosoma cruzi/efeitos dos fármacos
4.
J Appl Oral Sci ; 27: e20180699, 2019 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-31411265

RESUMO

OBJECTIVE: This study investigated the role of extracellular deoxyribonucleic acid (eDNA) on Enterococcus faecalis ( E. faecalis ) biofilm and the susceptibility of E. faecalis to sodium hypochlorite (NaOCl). METHODOLOGY: E. faecalis biofilm was formed in bovine tooth specimens and the biofilm was cultured with or without deoxyribonuclease (DNase), an inhibitor of eDNA. Then, the role of eDNA in E. faecalis growth and biofilm formation was investigated using colony forming unit (CFUs) counting, eDNA level assay, crystal violet staining, confocal laser scanning microscopy, and scanning electron microscopy. The susceptibility of E. faecalis biofilm to low (0.5%) or high (5%) NaOCl concentrations was also analyzed by CFU counting. RESULTS: CFUs and biofilm formation decreased significantly with DNase treatment (p<0.05). The microstructure of DNase-treated biofilms exhibited less structured features when compared to the control. The volume of exopolysaccharides in the DNase-treated biofilm was significantly lower than that of control (p<0.05). Moreover, the CFUs, eDNA level, biofilm formation, and exopolysaccharides volume were lower when the biofilm was treated with DNase de novo when compared to when DNase was applied to matured biofilm (p<0.05). E. faecalis in the biofilm was more susceptible to NaOCl when it was cultured with DNase (p<0.05). Furthermore, 0.5% NaOCl combined with DNase treatment was as efficient as 5% NaOCl alone regarding susceptibility (p>0.05). CONCLUSIONS: Inhibition of eDNA leads to decrease of E. faecalis biofilm formation and increase of susceptibility of E. faecalis to NaOCl even at low concentrations. Therefore, our results suggest that inhibition of eDNA would be beneficial in facilitating the efficacy of NaOCl and reducing its concentration.


Assuntos
Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , DNA Bacteriano/farmacologia , Desoxirribonucleases/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/crescimento & desenvolvimento , Hipoclorito de Sódio/farmacologia , Animais , Bovinos , Contagem de Colônia Microbiana , Cavidade Pulpar/microbiologia , Testes de Sensibilidade Microbiana , Microscopia Confocal , Microscopia Eletrônica de Varredura , Polissacarídeos Bacterianos/isolamento & purificação , Reprodutibilidade dos Testes , Fatores de Tempo
5.
Braz Oral Res ; 33: e057, 2019 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-31365704

RESUMO

The aim of this study was to evaluate the effect of peracetic acid (PAA) as a single irrigant on the smear layer, on the intraradicular dentinal bond strength, and on the penetrability of an epoxy-based resin sealer into the dentinal tubules. A total of 120 roots were distributed into 4 groups according to the irrigant used in root canal preparation: 1% PAA (PAA); 2.5% NaOCl followed by final irrigation with 17% EDTA and 2.5% NaOCl (NaOCl-EDTA-NaOCl); 2.5% NaOCl (NaOCl); and saline solution (SS). The smear layer was evaluated using scanning electron microscopy. The bond strength of an epoxy-based resin sealer (AH Plus) to root dentin was evaluated by the push-out test and penetrability of the sealer into dentinal tubules was observed by confocal laser microscopy. The results were analyzed by the Kruskal-Wallis and the Dunn post-test (α = 0.05). The use of 1% PAA as single root canal irrigant provided smear layer removal and improved the penetrability and bond strength of AH Plus to root dentin in a manner similar to that of the NaOCl-EDTA-NaOCl group (p > 0.05). The NaOCl and SS groups had higher values of smear layer and lower values of sealer penetrability and dentin bond strength than the PAA and NaOCl-EDTA-NaOCl groups (p < 0.05). Thus, 1% PAA has the potential to be used as a single irrigant in root canals.


Assuntos
Cavidade Pulpar/efeitos dos fármacos , Dentina/efeitos dos fármacos , Resinas Epóxi/química , Ácido Peracético/química , Materiais Restauradores do Canal Radicular/química , Irrigantes do Canal Radicular/química , Camada de Esfregaço/química , Colagem Dentária/métodos , Cavidade Pulpar/química , Dentina/química , Ácido Edético/química , Humanos , Teste de Materiais , Microscopia Confocal , Microscopia Eletrônica de Varredura , Valores de Referência , Reprodutibilidade dos Testes , Hipoclorito de Sódio/química
6.
Anticancer Res ; 39(8): 4533-4537, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31366556

RESUMO

BACKGROUND/AIM: Serum-derived macrophage activating factor, serum-MAF, is known to increase the phagocytic activity of macrophages by enhancing the engulfment efficiency. To elucidate the mechanisms underlying phagocytic activation, morphological changes were observed and analyzed. MATERIALS AND METHODS: Morphological changes in macrophages were observed and quantitatively analyzed using scanning electron microscope (SEM) and confocal microscope. RESULTS: SEM and confocal microscopy images revealed frill-like structures and active actin accumulations, respectively, in serum-MAF treated macrophages. Actin accumulation was induced within 5 min following serum-MAF treatment. CONCLUSION: Serum-MAF induced a rapid rearrangement of cytoskeletal actin and enhanced phagocytic activity. Findings of the current study may contribute to the development of techniques that facilitate activation of the human immune system, which in turn may be beneficial for cancer immunotherapy.


Assuntos
Actinas/química , Macrófagos/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-maf/farmacologia , Actinas/ultraestrutura , Humanos , Imunoterapia , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Microscopia Confocal , Microscopia Eletrônica de Varredura , Proteínas Proto-Oncogênicas c-maf/genética , Células U937 , Proteína de Ligação a Vitamina D/química , Proteína de Ligação a Vitamina D/metabolismo
7.
Cell Physiol Biochem ; 53(2): 337-354, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31373783

RESUMO

BACKGROUND/AIMS: The availability of truly maturated cardiomyocytic subtypes is a major prerequisite for cardiovascular cell replacement therapies. Pluripotent stem cells provide a suitable source for the development of new strategies to overcome enormous hurdles such as yield, purity and safety of in vitro generated cells. METHODS: To address these issues, we have refined existing forward programming protocols by combining forced exogenous overexpression of the early cardiovascular transcription factor Nkx2.5 with a αMHC-promoter-based antibiotic selection step. Additionally, we applied small molecules such as ascorbic acid to enhance cardiomyogenic differentiation efficiency. Subsequently, we evaluated the cell fate of the resulting cardiomyocytes on the mRNA as well as protein levels. The latter was performed using high-resolution confocal microscopy. Furthermore, we examined the response of the cells` beating activities to pharmacological substance administration. RESULTS: Our results reveal an apparent influence of Nkx2.5 on the cell fate of ESC-derived cardiomyocytes. Resulting single cells exhibit characteristics of early ventricular cardiomyocytes, such as sarcomeric marker expression, spontaneous beating frequency, and distinct L-type calcium channel occurrence. CONCLUSION: Therefore, we demonstrate cardiovascular subtype forward programming of ESCs using a combination of transcription factors along with small molecule administration. However, our findings also underline current assumptions, that a terminal maturation of PSC derived cardiomyocytes in vitro is still an unsolved problem which urgently needs to be addressed in the field.


Assuntos
Reprogramação Celular , Células-Tronco Embrionárias/metabolismo , Proteína Homeobox Nkx-2.5/metabolismo , Miócitos Cardíacos/metabolismo , Animais , Ácido Ascórbico/farmacologia , Diferenciação Celular/efeitos dos fármacos , Células-Tronco Embrionárias/citologia , Proteína Homeobox Nkx-2.5/antagonistas & inibidores , Proteína Homeobox Nkx-2.5/genética , Camundongos , Microscopia Confocal , Miócitos Cardíacos/citologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Verapamil/farmacologia
8.
Medicine (Baltimore) ; 98(35): e16589, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31464893

RESUMO

RATIONALE: Lipid deposition on the cornea without previous infection, inflammation, vascularization, or trauma is idiopathic lipid keratopathy. In vivo laser confocal microscopy (IVCM) and anterior segment optical coherence tomography (AS-OCT) are 2 noninvasive methods that can help identify the structures and morphological characteristics of the focus. PATIENT CONCERNS: A 63-year-old woman with ipsilateral corneal lipid deposits developing from a small white spot into a yellow-white superotemporal elliptic shape within a year. AS-OCT showed peripheral deep stromal deposits. IVCM showed hyper-reflective material with typical crystalline-like or needle-like structures in the superotemporal area. DIAGNOSIS: Idiopathic lipid degeneration. INTERVENTIONS: Topical steroids eye drops 3 times a day for a month and further consultation every 3 months. OUTCOMES: This patient of idiopathic lipid keratopathy was observed every 3 months and till now we have reviewed this patient twice. Topical steroids eye drops were only used during the first month. No further development was observed about the lesion and the patient's visual acuity remained good. CONCLUSION: IVCM and AS-OCT can help identify the characteristic crystalline-like or needle-like hyper-reflective material that could help diagnosis of idiopathic lipid degeneration.


Assuntos
Córnea/diagnóstico por imagem , Doenças da Córnea/tratamento farmacológico , Metabolismo dos Lipídeos , Esteroides/administração & dosagem , Administração Tópica , Córnea/metabolismo , Doenças da Córnea/diagnóstico por imagem , Doenças da Córnea/metabolismo , Feminino , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Microscopia Confocal , Pessoa de Meia-Idade , Esteroides/farmacologia , Tomografia de Coerência Óptica , Resultado do Tratamento , Acuidade Visual
9.
Exp Parasitol ; 204: 107731, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31374185

RESUMO

Neospora caninum is an obligate intracellular parasite related to cases of abortion and fertility impairment in cattle. The control of the parasite still lacks an effective protective strategy and the understanding of key mechanisms for host infection might be crucial for identification of specific targets. There are many proteins related to important mechanisms in the host cell infection cycle such as adhesion, invasion, proliferation and immune evasion. The surface proteins, especially SRS (Surface Antigen Glycoprotein - Related Sequences), have been demonstrated to have a pivotal role in the adhesion and invasion processes, making them potential anti-parasite targets. However, several predicted surface proteins were not described concerning their function and importance in the parasite life cycle. As such, a novel SRS protein, NcSRS57, was described. NcSRS57 antiserum was used to detect SRS proteins by immunofluorescence in parasites treated or not with phosphatidylinositol-specific phospholipase C (PI-PLC). The treatment with PI-PLC also allowed the identification of NcSRS29B and NcSRS29C, which were the most abundant SRS proteins in the soluble fraction. Our data indicated that SRS proteins in N. caninum shared a high level of sequence similarity and were susceptible to PI-PLC. In addition, the description of the SRS members, regarding abundance, function and immunogenicity will be useful in guiding specific methods to control the mechanism of adhesion and invasion mediated by these surface proteins.


Assuntos
Antígenos de Protozoários/metabolismo , Antígenos de Superfície/metabolismo , Neospora/química , Fosfoinositídeo Fosfolipase C/farmacologia , Proteínas de Protozoários/metabolismo , Animais , Antígenos de Protozoários/genética , Antígenos de Protozoários/imunologia , Antígenos de Superfície/genética , Antígenos de Superfície/imunologia , Cercopithecus aethiops , Clonagem Molecular , DNA de Protozoário/isolamento & purificação , Eletroforese em Gel Bidimensional , Eletroforese em Gel de Poliacrilamida , Soros Imunes/imunologia , Soros Imunes/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Confocal , Neospora/efeitos dos fármacos , Neospora/genética , Neospora/imunologia , Fosfoinositídeo Fosfolipase C/metabolismo , Proteínas de Protozoários/genética , Proteínas de Protozoários/imunologia , Espectrometria de Massas em Tandem , Fosfolipases Tipo C/metabolismo , Fosfolipases Tipo C/farmacologia , Células Vero
10.
Anal Chim Acta ; 1078: 101-111, 2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31358207

RESUMO

A series of polymers and metal ions have been observed to be useful in triggering aggregation-induced emission (AIE) and AIE enhancement (AIEE) of thiolated gold nanoclusters (AuNCs). However, peptide-induced AIEE of thiolated AuNCs and their applications in biosensors have rarely been investigated. In this study, we showed that positively charged peptides induced efficient AIEE of negatively charged glutathione-capped AuNCs (GSH-AuNCs) through electrostatic attraction. In contrast to GSH-AuNCs, polyarginine (polyArg), a cationic peptide, stimulated the AIEE of the GSH-AuNCs, resulting in a 3.5-fold luminescence enhancement, 10-fold enhancement in quantum yield, 8-nm blueshift in the luminescence maximum, and a 2.1-fold increase in the mean luminescence lifetime. Four different AIEE-based biosensors with excellent selectivity and acceptable sensitivity were fabricated using cationic peptides as an AIEE-active trigger and as a biorecognition element. A heparin biosensor with a limit of detection (LOD) of 3 nM was constructed by combining AG73 peptide-mediated AIEE of the GSH-AuNCs and the specific interaction of AG73 peptides with heparin macromolecules. The concentration of human trypsin was selectively detected at a concentration as low as 1 nM using an arginine-glycine repeat peptide as an enzymatic substrate and as an AIEE-active trigger. Alkaline phosphatase (ALP)-catalyzed dephosphorylation of phosphopeptides paired with the corresponding product-mediated AIEE of the GSH-AuNCs was used for ALP sensing with an LOD of 0.3 U L-1. A peptide consisting of a cyclic RGD unit and an AIEE-active unit was designed to synthesize RGD-modified GSH-AuNC aggregates that can target αvß3 integrin receptors. These AIEE-based sensors were practically applied for the quantitative determination of heparin in human plasma, trypsin in human urine, and ALP in human plasma as well as for luminescent imaging of αvß3 integrin-overexpressing HeLa cells.


Assuntos
Glutationa/química , Ouro/química , Nanopartículas Metálicas/química , Peptídeos Cíclicos/química , Fosfopeptídeos/química , Fosfatase Alcalina/química , Técnicas Biossensoriais/métodos , Linhagem Celular Tumoral , Heparina/análise , Humanos , Hidrólise , Integrina alfaVbeta3/metabolismo , Limite de Detecção , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Peptídeos Cíclicos/síntese química , Peptídeos Cíclicos/metabolismo , Espectrometria de Fluorescência/métodos , Eletricidade Estática , Tripsina/análise , Tripsina/química
11.
Anal Chim Acta ; 1078: 176-181, 2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31358217

RESUMO

Intracellular microRNA (miRNA) analysis in single cell is highly informative and offers valuable insights to its physiological and pathological state, but it must confront the pivotal challenge of gene probe delivery and conditional release. Herein, we report an assembled DNA mini-hexahedron (DMH) that can selectively package and protect miRNA probe, target-cell-specific delivery and release it based on the target sequence recognition for intracellular miRNA detection. In brief, the DMH is self-assembled from six single-stranded oligonucleotide strands through rational design, one of which containing AS1411 sequence for specific uptake. Two fluorescent dye labeled recognition strands are inserted into two DMH edges with quencher groups through partially complementary hybridization. We find that this DMH possesses great biocompatibility, good trans-membrane ability and are able to protect the gene cargo against enzymatic degradation and protein binding. Fluorescence restoration caused by the target-mediated competitive chain replacement reaction allows to simultaneous detection of two cancer-related intracellular miRNAs with little false-positive signal, providing a powerful tool to discriminate healthy normal cell and cancerous cell. Thus, the construct opens a new avenue to circumvent the challenges in gene delivery, specific delivery and intrinsic interferences resistance.


Assuntos
DNA de Cadeia Simples/química , MicroRNAs/análise , Aptâmeros de Nucleotídeos/química , Aptâmeros de Nucleotídeos/genética , Sequência de Bases , Linhagem Celular Tumoral , DNA de Cadeia Simples/genética , Portadores de Fármacos/química , Fluoresceínas/química , Fluorescência , Corantes Fluorescentes/química , Humanos , MicroRNAs/genética , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Hibridização de Ácido Nucleico , Oligodesoxirribonucleotídeos/química , Oligodesoxirribonucleotídeos/genética
12.
J Med Microbiol ; 68(9): 1353-1358, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31271350

RESUMO

Introduction. Candida auris is a pathogenic yeast that mainly affects immunosuppressed patients and those with implanted medical devices. This pathogen also displays elevated resistance to common antifungals and high survival and spreading capacities. Since no antifungal breakpoints have yet been defined for this pathogen, the data obtained here can be useful for further research concerning treatment or implementation of a prevention and disinfection protocol. Our aim was to study the antifungal resistance of C. auris to current antifungals in planktonic and sessile states. Using confocal laser scanning microscopy and viable biomass production, we demonstrated the ability of C. auris to develop a mature biofilm. We compared the minimal inhibitory concentration (MIC) and the minimal biofilm eradication concentration (MBEC) for the C. auris DSM 21092 strain plus two clinical isolates, and the results were compared with those obtained for Candida albicans and Candida parapsilosis, two species strongly linked to bloodstream infections and infections associated with biomaterials. We found that the clinical isolates of C. auris were resistant to fluconazole and sensitive to echinocandins and polyenes. The C. auris biofilms did not show susceptibility to any antifungal agent, showing MBECs that were up to 512-fold higher than the MICs. These findings highlight the importance of biofilm formation as a key factor underlying the resistance of this species to antifungals and suggest that the presence of implantable medical devices is one of the major risk factors in immunocompromised patients.


Assuntos
Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Candida/efeitos dos fármacos , Candida/isolamento & purificação , Candida albicans/efeitos dos fármacos , Candida parapsilosis/efeitos dos fármacos , Candidíase/microbiologia , Contagem de Colônia Microbiana , Farmacorresistência Fúngica , Equinocandinas/farmacologia , Fluconazol/farmacologia , Humanos , Testes de Sensibilidade Microbiana , Microscopia Confocal , Polienos/farmacologia
13.
Biol Res ; 52(1): 36, 2019 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-31300048

RESUMO

BACKGROUND: Recent evidences indicated that some local anaesthetic agents played a role in inhibiting the proliferation of cancer cells; Whether ropivacaine is able to promote apoptosis of hepatocellular carcinoma (HCC) cells is still unclear. The aim of this study was to investigate the effect of ropivacaine on the apoptosis of HCC cells. METHODS: In the present study, we treated the HCC cell lines, Bel7402 and HLE with ropivacaine. MTT, DAPI stain, trypan blue exclusion dye assay, flow cytometry, electron microscopy, computational simulation, laser confocal microscope, Western blotting, and enzyme activity analysis of caspase-3 were applied to detect the growth and apoptosis of HCC cells and to explore the role mechanism of ropivacaine. RESULTS: Ropivacaine was able to inhibit proliferation and promote apoptosis of HCC cells in a dose- and time-dependent manner. Ropivacaine also has a trait to inhibit the migration of HCC cells; ropivacaine damaged the mitochondria of HCC cells. The results also indicated that ropivacaine was able to interact with caspase-3, promote cytoplasmic caspase-3 migration into the nucleus, stimulate cleavage of caspase-3 and PARP-1, caspase-9 proteins, inhibit the expression of Bcl-2, promote expression of Apaf-1 and mitochondria release cytochrome C, and activate the activity of caspase-3. CONCLUSIONS: Ropivacaine has a novel role in promoting apoptosis of HCC cells; The role mechanism of ropivacaine maybe involve in damaging the function of mitochondria and activating the caspase-3 signalling pathway in HCC cells. Our findings provide novel insights into the local anaesthetic agents in the therapy of HCC patients.


Assuntos
Anestésicos Locais/farmacologia , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/patologia , Caspase 3/metabolismo , Neoplasias Hepáticas/patologia , Ropivacaina/farmacologia , Apoptose/fisiologia , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Citometria de Fluxo , Humanos , Neoplasias Hepáticas/metabolismo , Microscopia Confocal , Microscopia de Fluorescência , Mitocôndrias/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos
15.
Chem Commun (Camb) ; 55(65): 9629-9632, 2019 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-31353368

RESUMO

Excessive accumulation of reducing agents in the ER leads to a constitutively high UPR. And the co-function of GSH, Cys and HOCl in biological processes is not well understood. To address this, a TP probe, NPCC, was developed for monitoring reductive stress in the ER. It can also distinguish cancer cells from normal cells.


Assuntos
Cumarínicos/química , Estresse do Retículo Endoplasmático/fisiologia , Retículo Endoplasmático/metabolismo , Corantes Fluorescentes/química , Pirazóis/química , Animais , Cumarínicos/síntese química , Cisteína/química , Cisteína/metabolismo , Corantes Fluorescentes/síntese química , Glutationa/química , Glutationa/metabolismo , Cabras , Células HeLa , Humanos , Ácido Hipocloroso/química , Ácido Hipocloroso/metabolismo , Camundongos , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Oxirredução , Pirazóis/síntese química , Peixe-Zebra
16.
Stud Health Technol Inform ; 262: 376-379, 2019 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-31349246

RESUMO

Brain tumor surgery is a complex medical procedure. The extent of tumor resection is essential for postoperative recovery and survival. The neurosurgeon needs to regularly check the borderline between healthy and cancerous tissue. Currently, tissue samples are extracted via instantaneous sections and sent to a neuropathologist for examination. This process takes time and requires extensive human resources. Confocal laser endomicroscopy (CLE) allows visualization of cellular structures in vivo without the need for sample extraction. This paper presents a conceptual model which serves as the hypothetical basis for a planned intervention study to evaluate CLE with the case example of brain tumor surgery. Using current literature and expert interviews, the model has been tested. The expected improvement of the medical outcome could be confirmed. Regarding the socioeconomic impact of CLE, the literature is scarce. Expert interviews confirmed the hypotheses that costs and the workload for clinical staff might increase.


Assuntos
Neoplasias Encefálicas , Endoscopia , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/cirurgia , Humanos , Microscopia Confocal
17.
J Photochem Photobiol B ; 197: 111534, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31279897

RESUMO

In the search for developing a biomedicine based nanomaterial for therapeutic applications, here we described a new benign development of Photo-triggered Gold nanodots capped mesoporous silica nanoparticles Au@MSNs loaded with capsaicin (Cap) for photothermal therapy of cancer cells. Electron microscopic techniques (SEM and TEM) studies depict the anisotropic shape of Cap-Au@MSNs with mean size ≈110 nm. The successful amine functionalization and covalent interaction of Au nanodots on the mesoporous silica surface were confirmed from the results of FTIR, XPS and UV-vis spectral analyses, which directly indicates the composition of synthesized mesoporous silica surface. Additionally, Dynamic Light Scattering (DLS) revealed that synthesized cap-AuMSNs were stable with highly negatively charged. Cap-AuMSNs exhibited extraordinary in vitro antitumor activity against the tested twp thyroid cancer cell lines (i.e., FTC-133 and B-CPAP). 3-(4, 5-Dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay determined that capsaicin and Cap-AuMSNs conferred strong cytotoxicity against the FTC-133 and B-CPAP cell lines. Further, evaluation of the mechanism showed that anticancer activity was achieved by inducing apoptosis in thyroid cancer cells. In addition, we found that such compounds exhibited promising antimetastatic activity and reduced the invasiveness of cancer cells. Hence, we suggesting that these Cap-Au@MSNs can be used as promising candidates for cancer therapy and deserve further investigation.


Assuntos
Capsaicina/química , Raios Infravermelhos , Nanopartículas/química , Nanoestruturas/química , Dióxido de Silício/química , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Capsaicina/farmacologia , Capsaicina/uso terapêutico , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Ouro/química , Humanos , Microscopia Confocal , Fotoquimioterapia , Porosidade , Câncer Papilífero da Tireoide/tratamento farmacológico , Câncer Papilífero da Tireoide/patologia
18.
Chem Commun (Camb) ; 55(58): 8494-8497, 2019 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-31268095

RESUMO

A rational strategy was reported to construct boranil complexes (DPFB derivatives) with unique aggregation-induced emission effects by installing phenyl rings in the anil ligand as the intramolecular rotors. In view of the good biocompatibility and suitable lipophilicity, DPFB derivatives can serve as excellent fluorescent probes for specific imaging of lipid droplets in living cells and yolk lipids in zebrafish.


Assuntos
Compostos de Anilina/química , Compostos de Boro/química , Complexos de Coordenação/química , Corantes Fluorescentes/química , Gotículas Lipídicas/metabolismo , Compostos de Anilina/síntese química , Animais , Compostos de Boro/síntese química , Complexos de Coordenação/síntese química , Fluorescência , Corantes Fluorescentes/síntese química , Células HeLa , Humanos , Ligantes , Microscopia Confocal/métodos , Estrutura Molecular , Peixe-Zebra
19.
J Forensic Leg Med ; 66: 162-166, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31325687

RESUMO

In deaths due to electrocution intraepidermal separation, vacuolation of epidermal cells, "swiss cheese" aspect of the superficial part of epidermis (swollen and with multiple vescicles), metallization, necrosis of collagenous fibers, cardiomyocytes alterations are microscopically described. No cardiac nerve damage due to electricity is actually reported in literature. In this work we tried to find new morphological signs in the hearts of deaths due to electrocution. In three cases of deaths due to electrocution, in which forensic autopsies were performed within 36 h of the death, heart specimens were taken at the level of common trunk of the left coronary artery. The myocardium histological examination at optical microscope and Confocal Laser Scanning Microscope (CLSM) revealed fragmentation of cardiomyocytes, nerve trunks damage with hydropic swelling of the nerve fibers, interstitial and sub-nerve-sheath edema, very dishomogenous distribution of the natural fluorescence of the neurofilaments, coarctation of epicardial gangliar cells with cytoplasmic cleft and irregular fluorescence pattern. Identification of S-100 protein by immunohistochemistry can help to better observe the hydropic swelling of the nerve fibers and the central cytoplasmic clefts. These alterations could be used in future as specific signs of the passage of the electrical current through the heart. We recommend heart specimens at the level of common trunk of the left coronary artery in all the cases of suspected electrocution, to better evaluate cardiac nerve trunks damages and alterations.


Assuntos
Traumatismos por Eletricidade/patologia , Microscopia Confocal , Miocárdio/patologia , Adulto , Patologia Legal , Humanos , Imuno-Histoquímica , Masculino , Microscopia , Miócitos Cardíacos/patologia , Proteínas S100
20.
Food Chem ; 298: 125081, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31260952

RESUMO

The impact of kansui on the dough rheology and quality characteristics of chickpea-wheat composite flour-based noodles was investigated. For noodle dough, both the relative crystallinity and degree of order obtained from X-ray diffraction and Fourier transform infrared spectroscopy increased with the low level of kansui addition (0.5-1.5%), followed by a decreased trend at higher levels of alkali concentration (2.0-3.0%). Such variation was in accordance with the trend observed for G' and G″. Based on the confocal laser scanning microscopic observation for cooked noodle samples, the low concentration of kansui addition (0.5-1.5%) protected the well-embedded starch granules being disassociate from the gluten network, whereas with the increased alkali addition (2.0-3.0%), the protein network was impaired with a greater amount of ruptured starch granules being disassociated from the gluten network. The observation corresponded well with the electrophoretic profile, and the results of cooking properties, -SH content, and textural properties of cooked noodles.


Assuntos
Cicer/química , Farinha , Triticum/química , Carbonatos/química , Culinária , Qualidade dos Alimentos , Glutens/química , Microscopia Confocal , Potássio/química , Reologia , Espectroscopia de Infravermelho com Transformada de Fourier , Amido/química , Difração de Raios X
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